JPH0889232A - Apparatus for cell culture - Google Patents

Abstract

PURPOSE: To provide an apparatus for cell culture capable of sufficiently and homogeneously feeding oxygen to a cell without damaging the cell during the culture. CONSTITUTION: This apparatus for cell culture comprises a culture tank, a bubble feeder for feeding fine bubbles into the interior of the culture tank and a microvibration applicator (a piezoeletcric element 12, a vibrator, a diaphragm, etc.) installed facing to the interior of the culture tank).

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a culture device suitable for growing plant cells using a culture medium.

[0002]

2. Description of the Related Art There is already known a technique for industrially growing plant cells by culturing callus produced by dedifferentiation of plant cells in a liquid medium while agitating in a liquid medium. Although the technique of culturing plant tissue under such aerobic conditions is basically established, in the growth of cells,
There is a problem that cells are easily aggregated into lumps during culturing, and if the cells are strongly stirred to prevent it, the cells are easily damaged by mechanical stimulation of stirring. For this reason, various culture conditions have been studied in order to avoid such damage to cell tissues in culture and to bring the cells in culture into contact with a sufficient amount of air (oxygen).

For example, in the culture apparatus described in JP-A-2-142464 and JP-A-4-258285, a cylinder is provided inside the culture tank, and air bubbles are supplied toward the inside of the cylinder, and The culture fluid circulates inside and outside the cylinder in order to appropriately supply air to the cells and prevent damage to the cells.

Japanese Patent Laid-Open No. 4-248981 discloses
A stirring blade is provided in the culture tank, a culture container containing a culture solution and cells is installed in the stirring blade, and by rotating the culture container, a shaking action is given to the proliferating cells,
Disclosed is an invention adapted to prevent supply and aggregation of oxygen to cells.

On the other hand, studies have also been conducted on a device for supplying bubbles to a culture solution, and in Japanese Patent Laid-Open No. 5-292942, a sintered metal element is used as a device for supplying bubbles.
An invention is disclosed in which the supply rate of oxygen to the culture solution of the bubble supply device and the stirring blade is increased.

[0006]

DISCLOSURE OF THE INVENTION The present invention provides a cell culture device which enables sufficient oxygen supply to cells without damaging the cells in culture.

[0007]

[Means for Solving the Problems] The above-mentioned problems are caused by a culture tank,
The problem is solved by the cell culture device of the present invention, which comprises a bubble supply device for supplying fine bubbles to the inside of the culture tank and a microvibration applying means provided inside the culture tank. Examples of the microvibration applying means used in the present invention include a piezoelectric element, a diaphragm, a vibrator, and the like, and the piezoelectric element is particularly preferable.

The present invention will now be described in detail with reference to the accompanying drawings.

FIG. 1 shows an example of the configuration of the cell culture device of the present invention. That is, the cell culture device basically comprises a culture tank 2 for containing the culture medium 1 and a bubble supply device for supplying an oxygen-containing gas (usually air) as fine bubbles to the culture medium.

The air bubble supply device includes a pipe 5 equipped with an air pump 3 and a sterilization filter 4, a humidifier 7 containing sterile water 6 for humidifying the air supplied from the pipe 5, and the humidified air as air bubbles. Porous plate 8 supplied to culture solution 1
Consists of As the porous plate, those made of ceramic, glass, sintered metal, fluororesin, etc. are known, and metal filters are also known. As the porous plate used in the present invention, it is preferable to use a metal filter having high strength and low pressure loss.

The porous plate 8 is usually installed at the bottom or bottom of the culture tank 1. The air supplied to the culture solution 1 is
It is discharged to the outside through a condenser 10 from a pipe 9 provided at the top or the top of the culture tank. The culture device of FIG. 1 is provided with a stirring device 11 that uses mechanical rotation of stirring blades to stir the culture solution 1. However, in the cell culture device of the present invention, such mechanical stirring device 11 does not necessarily have to be installed. However,
It is preferable to install a stirring device in order to prevent cell aggregation.
Alternatively, the circulation means of the culture solution as disclosed in the above-mentioned publication may be used together.

The cell culture apparatus of FIG. 1 is further provided with a piezoelectric element 12 which is a typical example of a microvibration applying means which is a characteristic constituent feature of the present invention, and the piezoelectric element 12 is connected to an external power source 13. ing. The piezoelectric element 12 exhibits minute vibrations when driven by electrical energy, and the vibrations propagate to the culture medium 1, so that coalescence of bubbles supplied to the culture medium is effectively suppressed. Therefore, it becomes easy to uniformly supply oxygen to the cells, and the aggregation of the cells is suppressed accordingly.

A piezoelectric element, which is a typical example of the microvibration applying means used in the cell culture apparatus of the present invention, is known as an actuator element.
It is an element which itself exhibits minute vibrations. As the piezoelectric element, various types of configurations such as a unimorph type, a bimorph type, a lever type, and a scaled insect type are known, and any type can be used. Further, the microvibration applying means is not limited to the piezoelectric element, and is not particularly limited as long as it can apply microvibration to the culture solution introduced into the culture tank at a high frequency.
As examples other than the piezoelectric element, the above-mentioned diaphragm and vibrator can be cited.

In the cell culture apparatus of FIG. 1, the piezoelectric element 1
Although 2 is installed on the wall surface of the culture tank 2 so as to be in contact with the culture solution 1, the installation position of the piezoelectric element may be various places provided that it is in contact with the culture solution. For example,
It may be provided on the bottom of the culture tank 2, or the stirring device 11
You may attach to. Further, when an auxiliary means for facilitating the circulation and movement of the culture solution is provided in the stirring tank, such auxiliary means may be provided. There is no particular limitation on the number of microvibration applying means such as piezoelectric elements installed in the cell culture device of the present invention. The vibration can be changed as desired depending on the type of the means used, the size, the electric energy applied to drive the vibration, etc., so consider the type, size, culture conditions, etc. of the target cell culture device. The optimum piezoelectric element and its driving condition can be selected.

[0015]

EFFECTS OF THE INVENTION In the cell culture device of the present invention, by utilizing minute vibration by means of minute vibration imparting means such as a piezoelectric element attached to a culture tank, it is possible to prevent damage to cells being cultured in a culture solution. The coalescence of bubbles of oxygen-containing gas such as air supplied to the culture solution in the culture device can be effectively suppressed. Therefore, the supply of oxygen to the cells in the culture medium is highly uniform, and the control of the cell culture state becomes easy. Moreover, since coalescence of bubbles is suppressed, cell aggregation is also suppressed.

[Brief description of drawings]

FIG. 1 is a diagram showing an example of the configuration of a cell culture device of the present invention.

[Explanation of symbols]

 1 Culture Solution 2 Culture Tank 3 Air Pump 4 Disinfection Filter 5 Pipe 6 Sterile Water 7 Humidifier 8 Porous Plate 9 Pipe 10 Condenser 11 Stirrer 12 Piezoelectric Element 13 External Power Supply

Claims (3)

[Claims] 1. A cell culture device comprising a culture tank, a bubble supply device for supplying fine bubbles to the inside of the culture tank, and a microvibration applying means provided inside the culture tank. 2. The cell culture device according to claim 1, wherein the microvibration applying means is a piezoelectric element. 3. The cell culture device according to claim 1, further comprising a mechanical stirring device.